Characterizing the role of MSRA in pancreatic tumorigenesis

表征 MSRA 在胰腺肿瘤发生中的作用

• 批准号: 10658248
• 负责人: Iok In Christine Chio
• 金额: $ 46.62万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-17 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10658248
• 关键词: Acute; Adenosine; Amino Acids; Back; Biochemical; Biological; Biological Assay; Biology; CRISPR/Cas technology; Cell Survival; Cells; Chemicals; Collaborations; Communication; Complementary DNA; Complex; Cysteine; Data; Doxycycline; Enzymes; Genetic; Genomic Instability; Growth; In Vitro; Invaded; KRAS oncogenesis; KRAS2 gene; Knock-in; Knock-out; Maintenance; Malignant Neoplasms; Malignant neoplasm of pancreas; Medical center; Metabolic; Metabolic Pathway; Metabolism; Methionine; Mitochondria; Modeling; Molecular; Mouse Strains; Mus; Mutate; Nature; Neoplasm Metastasis; Normal tissue morphology; Oncogenes; Organoids; Outcome; Oxidation-Reduction; Oxidoreductase; Pancreas; Pancreatic Ductal Adenocarcinoma; Patients; Peptides; Periodicity; Post-Translational Protein Processing; Predisposition; Presbyterian Church; Primary Neoplasm; Prognosis; Protein Biosynthesis; Proteins; Proteomics; Pyruvate Kinase; Reactive Oxygen Species; Resistance; Respiration; Role; Signal Transduction; Site; Specimen; Structure; Sulfhydryl Compounds; Sulfur; Testing; Therapeutic; Therapeutically Targetable; Tissue Microarray; Tissues; Translations; Tumor Tissue; biochemical tools; cell behavior; cell injury; cell motility; chemoproteomics; clinically relevant; clinically significant; design; ectoATPase; extracellular; glucose metabolism; in vivo; inhibitor; macromolecule; methionine sulfoxide; methionine sulfoxide reductase; migration; mouse model; mutant; neoplastic cell; novel; organoid transplantation; oxidation; pancreatic ductal adenocarcinoma cell; pancreatic neoplasm; pancreatic tumorigenesis; pharmacologic; premalignant; protein expression; transplant model; tumor growth; tumorigenesis; ubiquitin isopeptidase; ubiquitin-protein ligase; wound healing

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDA) is an aggressive malignancy that remains largely incurable. The KRAS oncogene is mutated in 95% of PDAs and acts as a potent driver of PDA growth and maintenance. Oncogenic KRAS induces a profound rewiring of metabolic pathways that are essential for PDA growth. Interestingly, this rewiring generates metabolic by-products, including reactive oxygen species (ROS), that can potentially oxidize macromolecules, resulting in genomic instability and toxic cellular damage. However, in addition to these indiscriminate activities, ROS can also initiate redox signaling through oxidative post-translational modifications of proteinaceous cysteines, such that toggling between the reduced and oxidized states of cysteine constitutes a “redox switch” that can regulate the structure and/or function of a protein. Indeed, using our pancreatic organoid model (Cell, 2015), we previously showed that oncogenic KRAS promotes pancreatic tumorigenesis in part by regulating the cyclic reduction/oxidation of cysteines on select proteins involved in the translation machinery (Cell, 2016), thereby promoting protein synthesis (Cell, 2016), glucose metabolism, and tumor growth (Nature Communications, 2019). While cysteine is well-recognized for its susceptibility to oxidation, it is often not appreciated that methionine, the other sulfur-containing amino acid, can be readily oxidized to methionine sulfoxide, and reduced by the oxidoreductase, methionine sulfoxide reductase A (MSRA). Using a tissue microarray of patient-derived pancreatic tumors and adjacent normal tissues, we observed a stage-dependent decrease in MSRA protein expression in tumor tissues. Consistently, acute knockout of murine MSRA using CRISPR/Cas9 in premalignant pancreatic organoids provides a migratory advantage in vitro and in vivo. Thus, we hypothesize that redox signaling through oxidative post-translational modification of methionine residues (Met-oxPTM) represents an unexplored mechanism that supports pancreatic metastasis. Herein, we propose to i) define the mechanism regulating MSRA expression in PDA, ii) characterize the functional and clinical significance of oxidized methionine peptides in PDA metastasis, and iii) define and characterize targetable vulnerabilities created by Met-oxPTM in PDA cells.

项目摘要 胰腺导管腺癌（PDA）是一种侵袭性的恶性肿瘤，在很大程度上仍然无法治愈。述KRAS 癌基因在95%的PDA中发生突变，并作为PDA生长和维持的有力驱动因素。致癌 KRAS诱导对PDA生长至关重要的代谢途径的深刻重新布线。有趣的是， 重新连接产生代谢副产物，包括活性氧（ROS），可能会氧化 大分子，导致基因组不稳定性和毒性细胞损伤。不过除了这些 活性氧还可以通过氧化翻译后修饰启动氧化还原信号传导 蛋白质半胱氨酸，使得半胱氨酸的还原态和氧化态之间的切换构成 一种“氧化还原开关”，可以调节蛋白质的结构和/或功能。事实上，利用我们的胰腺 在类器官模型（Cell，2015）中，我们先前表明致癌KRAS促进胰腺肿瘤发生， 部分通过调节参与翻译的选择蛋白质上的半胱氨酸的循环还原/氧化 机械（Cell，2016），从而促进蛋白质合成（Cell，2016），葡萄糖代谢和肿瘤生长 （Nature Communications，2019）。虽然半胱氨酸因其对氧化的敏感性而被广泛认可，但它通常是 没有意识到另一种含硫氨基酸甲硫氨酸可以容易地氧化为甲硫氨酸 亚砜，并通过氧化还原酶甲硫氨酸亚砜还原酶A（MSRA）还原。使用组织 在患者来源的胰腺肿瘤和邻近正常组织的微阵列中，我们观察到阶段依赖性的 降低肿瘤组织中MSRA蛋白表达。一致地，使用以下方法急性敲除鼠MSRA： 癌前胰腺类器官中的CRISPR/Cas9在体外和体内提供了迁移优势。因此，在本发明中， 我们假设通过蛋氨酸的氧化翻译后修饰的氧化还原信号 Met-oxPTM代表了支持胰腺转移的未探索机制。 在此，我们建议i）定义调节PDA中MSRA表达的机制，ii）表征PDA中MSRA的表达， 氧化甲硫氨酸肽在PDA转移中的功能和临床意义，和iii）定义和 表征Met-oxPTM在PDA单元中创建的可定位漏洞。